#METABOLOMICS WORKBENCH JXL_20250726_183107 DATATRACK_ID:6217 STUDY_ID:ST004119 ANALYSIS_ID:AN006830 PROJECT_ID:PR002589 VERSION 1 CREATED_ON August 18, 2025, 6:53 pm #PROJECT PR:PROJECT_TITLE Bashaga Mongolian medicine anti-liver based on 'intestinal flora-tryptophan PR:PROJECT_TITLE metabolism-liver axis', Study on the mechanism of fiber action PR:PROJECT_SUMMARY Rhododendron molle (Blume) G. Don (abbreviated as RM), known as 'Bashaga' in PR:PROJECT_SUMMARY traditional Mongolian medicine, was investigated. Mongolian medicine believes PR:PROJECT_SUMMARY that liver fibrosis is caused by the interaction of 'liver-gut' between blood PR:PROJECT_SUMMARY and heat meridians. 'Bashaga' is the main Mongolian medicine treatment for liver PR:PROJECT_SUMMARY disease. There is still a lack of modern scientific research to clarify its PR:PROJECT_SUMMARY effectiveness. The source and mechanism of action restrict the clinical PR:PROJECT_SUMMARY development and rational utilization of 'Bashaga'. Preliminary Studies have PR:PROJECT_SUMMARY found that tryptophan metabolic pathway affects the occurrence and development PR:PROJECT_SUMMARY of liver fibrosis. Previous research has shown that Bashaga acts by regulating PR:PROJECT_SUMMARY the 'intestinal microbiota-tryptophan metabolism-liver axis'. This project PR:PROJECT_SUMMARY intends to use a CCL4 induced liver fibrosis rat model, to evaluate the effect PR:PROJECT_SUMMARY of 'Bashaga' by physiological and biochemical indexes such as pathology, liver PR:PROJECT_SUMMARY injury and inflammatory factors. To study the efficacy of Bahsaga to treat liver PR:PROJECT_SUMMARY fibrosis, the method of combining tryptophan-targeted metabolome with metagenome PR:PROJECT_SUMMARY was used. We explored its regulation of intestinal flora structure, tryptophan PR:PROJECT_SUMMARY metabolites and intestinal barrier by constructing the 'Intestinal PR:PROJECT_SUMMARY flora-tryptophan metabolism-phenotypic function index' to reveal the PR:PROJECT_SUMMARY anti-hepatic fibrosis effect of Bashaga. The purpose of this study is to clarify PR:PROJECT_SUMMARY the original source and clinical rationality of Bashaga guided by efficacy. PR:INSTITUTE Baotou Medical College PR:LAST_NAME Jiang PR:FIRST_NAME Xiaolei PR:ADDRESS No. 31, Jianshe Road, Donghe District, Baotou, Inner Mongolia, 014040, China PR:EMAIL jxl15147689596@163.com PR:PHONE 15147689596 #STUDY ST:STUDY_TITLE Bashaga Mongolian medicine anti-liver based on ' intestinal flora-tryptophan ST:STUDY_TITLE metabolism-liver axis ' Study on the mechanism of fiber action ST:STUDY_SUMMARY Rhododendron molle (Blume) G. Don (abbreviated as RM), known as 'Bashaga' in ST:STUDY_SUMMARY traditional Mongolian medicine, was investigated. Mongolian medicine believes ST:STUDY_SUMMARY that liver fibrosis is caused by the interaction of 'liver-gut' between blood ST:STUDY_SUMMARY and heat meridians. 'Bashaga' is the main Mongolian medicine treatment for liver ST:STUDY_SUMMARY disease. There is still a lack of modern scientific research to clarify its ST:STUDY_SUMMARY effectiveness. The source and mechanism of action restrict the clinical ST:STUDY_SUMMARY development and rational utilization 'Bashaga'. Preliminary Studies have found ST:STUDY_SUMMARY that tryptophan metabolic pathway affects the occurrence and development of ST:STUDY_SUMMARY liver fibrosis. Previous research has shown that Bashaga acts by regulating the ST:STUDY_SUMMARY 'intestinal microbiota-tryptophan metabolism-liver axis'. This project intends ST:STUDY_SUMMARY to use a CCL4 induced liver fibrosis rat model, to evaluate the effect of ST:STUDY_SUMMARY 'Bashaga' by physiological and biochemical indexes such as pathology, liver ST:STUDY_SUMMARY injury and inflammatory factors. To study the efficacy of Bahsaga to treat liver ST:STUDY_SUMMARY fibrosis, the method of combining tryptophan-targeted metabolome with metagenome ST:STUDY_SUMMARY was used. We explored its regulation of intestinal flora structure, tryptophan ST:STUDY_SUMMARY metabolites and intestinal barrier by constructing the 'Intestinal ST:STUDY_SUMMARY flora-tryptophan metabolism-phenotypic function index' to reveal the ST:STUDY_SUMMARY anti-hepatic fibrosis effect of Bashaga. The purpose of this study is to clarify ST:STUDY_SUMMARY the original source and clinical rationality of Bashaga guided by efficacy. ST:INSTITUTE Baotou Medical College ST:LAST_NAME Jiang ST:FIRST_NAME Xiaolei ST:ADDRESS No. 31, Jianshe Road, Donghe District, Baotou, Inner Mongolia, 014040, China ST:EMAIL jxl15147689596@163.com ST:PHONE 15147689596 #SUBJECT SU:SUBJECT_TYPE Mammal SU:SUBJECT_SPECIES Rattus norvegicus SU:TAXONOMY_ID 10116 SU:GENDER Male #SUBJECT_SAMPLE_FACTORS: SUBJECT(optional)[tab]SAMPLE[tab]FACTORS(NAME:VALUE pairs separated by |)[tab]Raw file names and additional sample data SUBJECT_SAMPLE_FACTORS CON_1 CON_1 Sample source:feces | Sample source:normal saline | Concentration:0.90% | Treatment:NA Duration=2 weeks; RAW_FILE_NAME(Moulding)=CON_1.RAW SUBJECT_SAMPLE_FACTORS CON_2 CON_2 Sample source:feces | Sample source:normal saline | Concentration:0.90% | Treatment:NA Duration=2 weeks; RAW_FILE_NAME(Moulding)=CON_2.RAW SUBJECT_SAMPLE_FACTORS CON_3 CON_3 Sample source:feces | Sample source:normal saline | Concentration:0.90% | Treatment:NA Duration=2 weeks; RAW_FILE_NAME(Moulding)=CON_3.RAW SUBJECT_SAMPLE_FACTORS CON_4 CON_4 Sample source:feces | Sample source:normal saline | Concentration:0.90% | Treatment:NA Duration=2 weeks; RAW_FILE_NAME(Moulding)=CON_4.RAW SUBJECT_SAMPLE_FACTORS CON_5 CON_5 Sample source:feces | Sample source:normal saline | Concentration:0.90% | Treatment:NA Duration=2 weeks; RAW_FILE_NAME(Moulding)=CON_5.RAW SUBJECT_SAMPLE_FACTORS CON_6 CON_6 Sample source:feces | Sample source:normal saline | Concentration:0.90% | Treatment:NA Duration=2 weeks; RAW_FILE_NAME(Moulding)=CON_6.RAW SUBJECT_SAMPLE_FACTORS MOD_1 MOD_1 Sample source:feces | Sample source:normal saline | Concentration:0.90% | Treatment:40% CCl4 + corn oil（Two times each week） Duration=2 weeks; RAW_FILE_NAME(Moulding)=MOD_1.RAW SUBJECT_SAMPLE_FACTORS MOD_2 MOD_2 Sample source:feces | Sample source:normal saline | Concentration:0.90% | Treatment:40% CCl4 + corn oil（Two times each week） Duration=2 weeks; RAW_FILE_NAME(Moulding)=MOD_2.RAW SUBJECT_SAMPLE_FACTORS MOD_3 MOD_3 Sample source:feces | Sample source:normal saline | Concentration:0.90% | Treatment:40% CCl4 + corn oil（Two times each week） Duration=2 weeks; RAW_FILE_NAME(Moulding)=MOD_3.RAW SUBJECT_SAMPLE_FACTORS MOD_4 MOD_4 Sample source:feces | Sample source:normal saline | Concentration:0.90% | Treatment:40% CCl4 + corn oil（Two times each week） Duration=2 weeks; RAW_FILE_NAME(Moulding)=MOD_4.RAW SUBJECT_SAMPLE_FACTORS MOD_5 MOD_5 Sample source:feces | Sample source:normal saline | Concentration:0.90% | Treatment:40% CCl4 + corn oil（Two times each week） Duration=2 weeks; RAW_FILE_NAME(Moulding)=MOD_5.RAW SUBJECT_SAMPLE_FACTORS MOD_6 MOD_6 Sample source:feces | Sample source:normal saline | Concentration:0.90% | Treatment:40% CCl4 + corn oil（Two times each week） Duration=2 weeks; RAW_FILE_NAME(Moulding)=MOD_6.RAW SUBJECT_SAMPLE_FACTORS Sily_1 Sily_1 Sample source:feces | Sample source:Sily | Concentration:100 mg/kg (body weight) | Treatment:40% CCl4 + corn oil（Two times each week） Duration=2 weeks; RAW_FILE_NAME(Moulding)=Sily_1.RAW SUBJECT_SAMPLE_FACTORS Sily_2 Sily_2 Sample source:feces | Sample source:Sily | Concentration:100 mg/kg (body weight) | Treatment:40% CCl4 + corn oil（Two times each week） Duration=2 weeks; RAW_FILE_NAME(Moulding)=Sily_2.RAW SUBJECT_SAMPLE_FACTORS Sily_3 Sily_3 Sample source:feces | Sample source:Sily | Concentration:100 mg/kg (body weight) | Treatment:40% CCl4 + corn oil（Two times each week） Duration=2 weeks; RAW_FILE_NAME(Moulding)=Sily_3.RAW SUBJECT_SAMPLE_FACTORS Sily_4 Sily_4 Sample source:feces | Sample source:Sily | Concentration:100 mg/kg (body weight) | Treatment:40% CCl4 + corn oil（Two times each week） Duration=2 weeks; RAW_FILE_NAME(Moulding)=Sily_4.RAW SUBJECT_SAMPLE_FACTORS Sily_5 Sily_5 Sample source:feces | Sample source:Sily | Concentration:100 mg/kg (body weight) | Treatment:40% CCl4 + corn oil（Two times each week） Duration=2 weeks; RAW_FILE_NAME(Moulding)=Sily_5.RAW SUBJECT_SAMPLE_FACTORS Sily_6 Sily_6 Sample source:feces | Sample source:Sily | Concentration:100 mg/kg (body weight) | Treatment:40% CCl4 + corn oil（Two times each week） Duration=2 weeks; RAW_FILE_NAME(Moulding)=Sily_6.RAW SUBJECT_SAMPLE_FACTORS RMH_1 RMH_1 Sample source:feces | Sample source:RM | Concentration:0.315g/kg (body weight) | Treatment:40% CCl4 + corn oil（Two times each week） Duration=2 weeks; RAW_FILE_NAME(Moulding)=RMH_1.RAW SUBJECT_SAMPLE_FACTORS RMH_2 RMH_2 Sample source:feces | Sample source:RM | Concentration:0.315g/kg (body weight) | Treatment:40% CCl4 + corn oil（Two times each week） Duration=2 weeks; RAW_FILE_NAME(Moulding)=RMH_2.RAW SUBJECT_SAMPLE_FACTORS RMH_3 RMH_3 Sample source:feces | Sample source:RM | Concentration:0.315g/kg (body weight) | Treatment:40% CCl4 + corn oil（Two times each week） Duration=2 weeks; RAW_FILE_NAME(Moulding)=RMH_3.RAW SUBJECT_SAMPLE_FACTORS RMH_4 RMH_4 Sample source:feces | Sample source:RM | Concentration:0.315g/kg (body weight) | Treatment:40% CCl4 + corn oil（Two times each week） Duration=2 weeks; RAW_FILE_NAME(Moulding)=RMH_4.RAW SUBJECT_SAMPLE_FACTORS RMH_5 RMH_5 Sample source:feces | Sample source:RM | Concentration:0.315g/kg (body weight) | Treatment:40% CCl4 + corn oil（Two times each week） Duration=2 weeks; RAW_FILE_NAME(Moulding)=RMH_5.RAW SUBJECT_SAMPLE_FACTORS RMH_6 RMH_6 Sample source:feces | Sample source:RM | Concentration:0.315g/kg (body weight) | Treatment:40% CCl4 + corn oil（Two times each week） Duration=2 weeks; RAW_FILE_NAME(Moulding)=RMH_6.RAW SUBJECT_SAMPLE_FACTORS RML_1 RML_1 Sample source:feces | Sample source:RM | Concentration:0.07875g/kg (body weight) | Treatment:40% CCl4 + corn oil（Two times each week） Duration=2 weeks; RAW_FILE_NAME(Moulding)=RML_1.RAW SUBJECT_SAMPLE_FACTORS RML_2 RML_2 Sample source:feces | Sample source:RM | Concentration:0.07875g/kg (body weight) | Treatment:40% CCl4 + corn oil（Two times each week） Duration=2 weeks; RAW_FILE_NAME(Moulding)=RML_2.RAW SUBJECT_SAMPLE_FACTORS RML_3 RML_3 Sample source:feces | Sample source:RM | Concentration:0.07875g/kg (body weight) | Treatment:40% CCl4 + corn oil（Two times each week） Duration=2 weeks; RAW_FILE_NAME(Moulding)=RML_3.RAW SUBJECT_SAMPLE_FACTORS RML_4 RML_4 Sample source:feces | Sample source:RM | Concentration:0.07875g/kg (body weight) | Treatment:40% CCl4 + corn oil（Two times each week） Duration=2 weeks; RAW_FILE_NAME(Moulding)=RML_4.RAW SUBJECT_SAMPLE_FACTORS RML_5 RML_5 Sample source:feces | Sample source:RM | Concentration:0.07875g/kg (body weight) | Treatment:40% CCl4 + corn oil（Two times each week） Duration=2 weeks; RAW_FILE_NAME(Moulding)=RML_5.RAW SUBJECT_SAMPLE_FACTORS RML_6 RML_6 Sample source:feces | Sample source:RM | Concentration:0.07875g/kg (body weight) | Treatment:40% CCl4 + corn oil（Two times each week） Duration=2 weeks; RAW_FILE_NAME(Moulding)=RML_6.RAW SUBJECT_SAMPLE_FACTORS QC_1 QC_1 Sample source:feces | Sample source:NA | Concentration:NA | Treatment:NA Duration=2 weeks; RAW_FILE_NAME(Moulding)=QC_1.RAW SUBJECT_SAMPLE_FACTORS QC_2 QC_2 Sample source:feces | Sample source:NA | Concentration:NA | Treatment:NA Duration=2 weeks; RAW_FILE_NAME(Moulding)=QC_2.RAW SUBJECT_SAMPLE_FACTORS QC_3 QC_3 Sample source:feces | Sample source:NA | Concentration:NA | Treatment:NA Duration=2 weeks; RAW_FILE_NAME(Moulding)=QC_3.RAW SUBJECT_SAMPLE_FACTORS QC_4 QC_4 Sample source:feces | Sample source:NA | Concentration:NA | Treatment:NA Duration=2 weeks; RAW_FILE_NAME(Moulding)=QC_4.RAW SUBJECT_SAMPLE_FACTORS QC_5 QC_5 Sample source:feces | Sample source:NA | Concentration:NA | Treatment:NA Duration=2 weeks; RAW_FILE_NAME(Moulding)=QC_5.RAW SUBJECT_SAMPLE_FACTORS QC_6 QC_6 Sample source:feces | Sample source:NA | Concentration:NA | Treatment:NA Duration=2 weeks; RAW_FILE_NAME(Moulding)=QC_6.RAW SUBJECT_SAMPLE_FACTORS QC_7 QC_7 Sample source:feces | Sample source:NA | Concentration:NA | Treatment:NA Duration=2 weeks; RAW_FILE_NAME(Moulding)=QC_7.RAW SUBJECT_SAMPLE_FACTORS QC_8 QC_8 Sample source:feces | Sample source:NA | Concentration:NA | Treatment:NA Duration=2 weeks; RAW_FILE_NAME(Moulding)=QC_8.RAW #COLLECTION CO:COLLECTION_SUMMARY Fresh feces were collected by pressing method before dissection of rats. CO:SAMPLE_TYPE Feces CO:STORAGE_CONDITIONS -80℃ #TREATMENT TR:TREATMENT_SUMMARY Rats were treated with an extract of Rhododendron molle (Blume) G. Don (RM), TR:TREATMENT_SUMMARY silymarin (Sily) as positive control and normal saline as control. #SAMPLEPREP SP:SAMPLEPREP_SUMMARY After the fecal sample was taken out from the refrigerator at − 80 ° C, 100 SP:SAMPLEPREP_SUMMARY mg of feces was taken, added with 1000 uL of 75 % methanol in a 1.5 mL SP:SAMPLEPREP_SUMMARY centrifuge tube, homogenized, suspended for 30 s, and sonicated for 10 min. SP:SAMPLEPREP_SUMMARY After centrifugation at 13,000 rpm, the supernatant was filtered through the SP:SAMPLEPREP_SUMMARY membrane, and the first three drops were not used as samples for fecal analysis. #CHROMATOGRAPHY CH:CHROMATOGRAPHY_TYPE Reversed phase CH:INSTRUMENT_NAME Waters Acquity I-Class CH:COLUMN_NAME Waters ACQUITY UPLC HSS T3 (100 x 2.1mm, 1.8um) CH:SOLVENT_A 100% Acetonitrile; 0.1 % formic acid CH:SOLVENT_B 100% Water; 0.1 % formic acid CH:FLOW_GRADIENT 0-4min，1%-50% A；4-6min：50%-90% A，6-7min：90%-100%A，7-8：100%A CH:FLOW_RATE 5 microLiters/min CH:COLUMN_TEMPERATURE 30℃ CH:CHROMATOGRAPHY_COMMENTS ACQUITY UPLC HSS T3 ( 2.1× 100 mm，1. 8 μm) #ANALYSIS AN:ANALYSIS_TYPE MS #MS MS:INSTRUMENT_NAME Thermo Q Exactive Orbitrap MS:INSTRUMENT_TYPE Orbitrap MS:MS_TYPE ESI MS:ION_MODE POSITIVE MS:MS_COMMENTS Progenesis QI software can be used to analyze the original data of metabolomics. MS:MS_COMMENTS The software can normalize the intensity of each ion relative to the total ion MS:MS_COMMENTS number, and generate a data matrix, which includes retention time, MS:MS_COMMENTS mass-to-charge ratio and normalized peak area, so as to obtain more insights and MS:MS_COMMENTS useful results. The data matrix was subjected to unsupervised principal MS:MS_COMMENTS component analysis ( PCA ) and supervised partial least squares-discriminant MS:MS_COMMENTS analysis ( PLS-DA ) using EZinfor software. #MS_METABOLITE_DATA MS_METABOLITE_DATA:UNITS normalized peak area MS_METABOLITE_DATA_START Samples CON_1 CON_2 CON_3 CON_4 CON_5 CON_6 MOD_1 MOD_2 MOD_3 MOD_4 MOD_5 MOD_6 RML_1 RML_2 RML_3 RML_4 RML_5 RML_6 RMH_1 RMH_2 RMH_3 RMH_4 RMH_5 RMH_6 Factors Sample source:feces | Sample source:normal saline | Concentration:0.90% | Treatment:NA Sample source:feces | Sample source:normal saline | Concentration:0.90% | Treatment:NA Sample source:feces | Sample source:normal saline | Concentration:0.90% | Treatment:NA Sample source:feces | Sample source:normal saline | Concentration:0.90% | Treatment:NA Sample source:feces | Sample source:normal saline | Concentration:0.90% | Treatment:NA Sample source:feces | Sample source:normal saline | Concentration:0.90% | Treatment:NA Sample source:feces | Sample source:normal saline | Concentration:0.90% | Treatment:40% CCl4 + corn oil（Two times each week） Sample source:feces | Sample source:normal saline | Concentration:0.90% | Treatment:40% CCl4 + corn oil（Two times each week） Sample source:feces | Sample source:normal saline | Concentration:0.90% | Treatment:40% CCl4 + corn oil（Two times each week） Sample source:feces | Sample source:normal saline | Concentration:0.90% | Treatment:40% CCl4 + corn oil（Two times each week） Sample source:feces | Sample source:normal saline | Concentration:0.90% | Treatment:40% CCl4 + corn oil（Two times each week） Sample source:feces | Sample source:normal saline | Concentration:0.90% | Treatment:40% CCl4 + corn oil（Two times each week） Sample source:feces | Sample source:RM | Concentration:0.07875g/kg (body weight) | Treatment:40% CCl4 + corn oil（Two times each week） Sample source:feces | Sample source:RM | Concentration:0.07875g/kg (body weight) | Treatment:40% CCl4 + corn oil（Two times each week） Sample source:feces | Sample source:RM | Concentration:0.07875g/kg (body weight) | Treatment:40% CCl4 + corn oil（Two times each week） Sample source:feces | Sample source:RM | Concentration:0.07875g/kg (body weight) | Treatment:40% CCl4 + corn oil（Two times each week） Sample source:feces | Sample source:RM | Concentration:0.07875g/kg (body weight) | Treatment:40% CCl4 + corn oil（Two times each week） Sample source:feces | Sample source:RM | Concentration:0.07875g/kg (body weight) | Treatment:40% CCl4 + corn oil（Two times each week） Sample source:feces | Sample source:RM | Concentration:0.315g/kg (body weight) | Treatment:40% CCl4 + corn oil（Two times each week） Sample source:feces | Sample source:RM | Concentration:0.315g/kg (body weight) | Treatment:40% CCl4 + corn oil（Two times each week） Sample source:feces | Sample source:RM | Concentration:0.315g/kg (body weight) | Treatment:40% CCl4 + corn oil（Two times each week） Sample source:feces | Sample source:RM | Concentration:0.315g/kg (body weight) | Treatment:40% CCl4 + corn oil（Two times each week） Sample source:feces | Sample source:RM | Concentration:0.315g/kg (body weight) | Treatment:40% CCl4 + corn oil（Two times each week） Sample source:feces | Sample source:RM | Concentration:0.315g/kg (body weight) | Treatment:40% CCl4 + corn oil（Two times each week） Niacinamide 1126.197649 1003.426914 1278.482446 1469.873797 708.438843 750.0401355 83600.8249 77132.38635 71732.0438 67198.72006 166734.6758 67549.55381 14171.64065 20582.088 18373.91664 16146.47416 12147.00846 13328.63504 95488.02621 118069.3604 330052.7609 298562.8277 224618.6138 64816.33589 Pregnanediol 3-O-glucuronide 4829.500744 6766.625949 7829.670022 10914.53759 8344.287365 9936.435538 10595.56601 10623.18615 10274.73423 8338.291591 9196.981106 15482.09159 26644.4258 19035.77021 23378.84081 24913.79613 35505.78175 26210.13198 87585.36769 100483.3372 292335.8017 271646.6745 229222.6236 59582.72065 Saccharopine 51126.79494 58478.03423 54846.39733 61050.24903 53201.20668 35354.4016 3397.090278 8851.822221 4935.566234 5286.848921 5183.645041 3064.807817 2262.096809 1874.761181 739.2781185 1666.984231 1621.456987 1319.61154 91170.48734 101581.6202 311357.488 280526.8208 199387.9713 53649.67276 Stachyose 0 0 0 0 0 0 230145.7357 107019.1359 86393.66518 49575.44122 106529.1215 90511.38157 6293.208013 7979.238309 9329.872874 10481.28782 6343.951652 14506.51441 47547.27756 50912.69686 117288.8641 89619.68557 88485.72147 16672.11819 MS_METABOLITE_DATA_END #METABOLITES METABOLITES_START metabolite_name retention index  quantified m/z  PubChem ID KEGG Others Niacinamide 894278.466 123.055 936 C00153 M+H Pregnanediol 3-O-glucuronide 842564.7323 495.296 443486 — M+H Saccharopine 842209.1363 264.144 439207 C00449 M+H Stachyose 834899.317 665.215 43 C01620 M+H METABOLITES_END #END